REGULATION OF VIRUS INDUCED PROGRAMMED CELL DEATH

程序性细胞死亡或凋亡是一个内在的、由信号诱导的过程 细胞通过这种方式自毁。这是一种提供控制的机制 细胞数量，因此对正常发育和 组织动态平衡。在人类中，异常的程序性细胞死亡是 与肿瘤发生、神经退行性疾病有关， 免疫缺陷和病毒致病机制。尽管细胞凋亡途径 在不同的有机体之间高度保守，并涉及中心性死亡 调节器，参与执行和调节的细胞机制 在很大程度上是未知的。诱导细胞凋亡的动物病毒 可以通过表达新的凋亡抑制因子来阻止细胞死亡。 因此，病毒干预宿主细胞凋亡的机制 提供了对细胞死亡程序的关键见解。这是一个长期的问题 这项提议的目的是定义分子机制 细胞凋亡是通过研究杆状病毒编码的 细胞凋亡调节因子、p35和IAP。我们的方法侧重于使用 杆状病毒感染的昆虫细胞是一种强大而又方便的系统 用于诱导和抑制细胞凋亡的分子分析。在……里面 建立在最新进展基础上的一系列综合实验 程序性细胞死亡，我们使用生物化学、遗传学和细胞生物学 P35和VIAP抗肿瘤分子机制的研究进展 细胞凋亡活性。功能丧失p35的广泛集合 突变将被用来描述分子间的相互作用 P35和CED-3/ICE相关的死亡蛋白，其中P35是一个有效的 抑制剂。通过对无脊椎动物CED-3/ICE样蛋白酶的研究(S) 在杆状病毒感染诱导下，我们还探讨了其作用机制。 参与激活和抑制这些关键的细胞死亡 酵素。在遗传学研究中，我们使用新的p35和iap表达 杆状病毒重组体和稳定表达的昆虫细胞 确定P35和VIAP的抗细胞凋亡活性 进入保守的死亡途径。最后，我们使用p35和iap作为工具 明确大型DNA病毒引发细胞凋亡的机制 它们的宿主细胞和致病作用。总而言之，这些 预计研究将通过以下方式对这些机制产生重要的洞察 其中程序性细胞死亡是受调控的，从而促进了长时间的 治疗细胞凋亡的新治疗策略的术语设计 相关的人类疾病。

Programmed cell death or apoptosis is a built-in, signal-induced process by which a cell self-destructs. It is a mechanism that provides control of cell numbers, and is therefore critical for normal development and tissue homeostasis. In humans, aberrant programmed cell death is associated with tumorigenesis, neurodegenerative diseases, immunodeficiency, and viral pathogenesis. Although the apoptotic pathway is highly conserved between diverse organisms and involves central death regulators, the cellular mechanisms involved in execution and regulation of apoptosis are largely unknown. Animal viruses that induce apoptosis can block cell death by the expression of novel apoptotic suppressors. The mechanisms for viral intervention of host apoptosis have therefore provided key insight into the cell death program. It is the long term objective of this proposal to define the molecular mechanisms by which apoptosis is regulated through the study of the baculovirus-encoded apoptotic regulators, p35 and iap. Our approach focuses on the use of baculovirus-infected insect cells as a powerful, yet convenient system for molecular analysis of the induction and suppression of apoptosis. In an integrated set of experiments that build on recent advances in programmed cell death, we use biochemical, genetic, and cell biology approaches to determine the molecular mechanism of P35 and vIAP anti- apoptotic activity. An extensive collection of loss-of-function p35 mutations will be used to characterize the molecular interaction between P35 and the CED-3/ICE-related death proteases, for which P35 is a potent inhibitor. By focusing on the invertebrate CED-3/ICE-like protease(s) induced by baculovirus infection, we also investigate the mechanisms involved in activation and inhibition of these critical cell death enzymes. In genetic studies, we use novel p35- and iap-expressing baculovirus recombinants, as well as stably transfected insect cell lines, to definitively place the anti-apoptotic activity of P35 and vIAP into the conserved death pathway. Lastly, we use p35 and iap as tools to define the mechanisms by which large DNA viruses trigger apoptosis in their host cell and contribute to pathogenesis. Collectively, these studies are expected to yield important insight into the mechanisms by which programmed cell death is regulated and thereby facilitate the long term design of novel therapeutic strategies for treatment of apoptosis- linked human disorders.